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Double-labelling methods used to diagnose apoptotic and necrotic cell degradations in copepod nauplii
Authors:S.?A.?Poulet  author-information"  >  author-information__contact u-icon-before"  >  mailto:poulet@sb-roscoff.fr"   title="  poulet@sb-roscoff.fr"   itemprop="  email"   data-track="  click"   data-track-action="  Email author"   data-track-label="  "  >Email author,M.?Richer de Forges,A.?Cueff,J.-F.?Lennon
Affiliation:(1) Station Biologique de Roscoff, CNRS/INSU/UPMC, BP 74, 29682 Roscoff, France
Abstract:Two double-labelling methods, Tunel+propidium iodide and Annexin V-FITC+propidium iodide, have been tested to diagnose cell degradation processes in N1–N2 nauplius stages produced by spawning females of the copepod Calanus helgolandicus fed either non-toxic (PRO: control) or toxic (TR1) diets under laboratory conditions. Observation of labelled samples with a confocal laser-scanning microscope revealed that the maternal-food effect, following absorption of the toxic diet, induced cell apoptosis and necrosis in 80–100% of the offspring, which sooner or later died. Similar cell damages were also observed in 18–50% of the nauplii produced by females incubated in filtered seawater. Such cell degradation processes and high mortality were not observed in nauplii produced by females fed the non-toxic diet. Our protocols allowed detection of cell degradations before death of unhealthy larvae, preferentially with a confocal microscope or a conventional visible-fluorescent-light microscope.Communicated by O. Kinne, Oldendorf/Luhe
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